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Klenow Fragment

  1. Nov 25, 2003 #1
    I know what it is, but my question is: What are some (specifically) popular applications? I am guessing some sort of specialized PCR??? Any thoughts would be appreciated.
     
  2. jcsd
  3. Nov 25, 2003 #2

    iansmith

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    I know we use it to double strand DNA for electrophoretic mobility shift assay (EMSA) and DNA footprintings.

    From the following site
    http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/enzymes/klenow.html

    1. Synthesis of double-stranded DNA from single-stranded templates
    2. Filling in recessed 3' ends of DNA fragments
    3. Digesting away protruding 3' overhangs
    4. Preparation of radioactive DNA probes
     
  4. Nov 25, 2003 #3
    Thank you Ian, that is just what I wanted to know!! :smile: This brings me to my next question I guess, being, what advantage would there be in utilizing JUST klenow fragment in synthesis of DS DNA fragments from SS DNA versus DNA polymerase III?
     
  5. Nov 25, 2003 #4

    iansmith

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    Klenow fragment can start synthesis from a primed single strand whereas DNA Pol III requires ssb protein in addition to a primed ss.
     
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